响应面法优化枯草芽孢杆菌产脂肪酶的合成培养基

doi:Q815

中国生物工程杂志

2010, Vol. 30

Issue (08): 100-105 DOI: Q815

技术与方法

响应面法优化枯草芽孢杆菌产脂肪酶的合成培养基

宋萍,戚小灵,胡燚,谢宁昌

南京工业大学生物与制药工程学院南京 210009

Optimization of Lipase Production Conditions by Bacillus subtilis Using Surface Methodology

SONG Ping,QI Xiao-ling,HU Yi,XIE Ning-chang

College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, China

全文: PDF(1485 KB) HTML

摘要：

对枯草芽孢杆菌(Bacillus subtilis) CICC 20034利用合成培养基液体发酵产脂肪酶的条件进行了优化。首先采用单因子实验筛选出最适诱导剂为三丁酸甘油酯，氮源为尿素，碳源为葡萄糖，无机盐为MgSO4。在此基础上，利用Plackett-Burman设计对影响产酶因素的效应进行评价，筛选出具有显著效应的三丁酸甘油酯、尿素、KH2PO4和培养基起始pH值四个最显著的因素。用最陡爬坡路径逼近最大产酶区域后，利用响应面中心组合设计对显著因素进行优化，获得最适合成培养基组分为：葡萄糖8 g/l，尿素8.57 g/l，三丁酸甘油酯 2.62 %，KH2PO4 2.59 g/l，MgSO4·7H2O 0.5 g/l，Tritonx-100 0.5 g/l，pH9.47。优化后的B. subtilis CICC 20034胞外脂肪酶活力达0.483 U /ml，比初始酶活力0.072 U /ml提高了6.7倍。

关键词： 脂肪酶; 枯草芽孢杆菌; Plackett-Burman设计; 响应面法; 优化

Abstract:

The fermentation conditions of lipase production by Bacillus subtilis CICC 20034 were optimized. Initially, the most suitable intruder tributyrin, nitrogen source urea, carbon glucose, and MgSO4 were selected according to single factorial experiments respectively. Based on the result, screening methodology Plackett-Burman design was used to evaluate the effects of twelve factors related to lipase production and four statistically significant factors tributyrin, urea, KH2PO4 and pH were selected. The path of steepest ascent was used to approach the optimal region of lipase production subsequently. Then, the optimal combined concentration for maximum enzyme activity were further optimized by response surface methodology and determined as follows: tributyrin 2. 62%, urea 8.57g/L, KH2PO4 2.59g/L and pH 9.47. The optimization of culture conditions of B.subtilis CICC 20034 led to a 6.7-fold increase in lipase production relative to initial result 0.072 /ml, which indicate that single factor in combination with response surface methodology, is an effective method for optimization of lipase production conditions by B.subtilis CICC 20034.

Key words: Lipase Bacillus subtilis Plackett-Burman design Response surface methodology Optimization

收稿日期: 2010-01-21 出版日期: 2010-08-25

基金资助:

国家自然科学基金重点项目(20936002）、国家自然科学基金 (20906049)资助项目

通讯作者: 谢宁昌 E-mail: biotech@njut.edu.cn

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引用本文:

宋萍戚小灵胡燚谢宁昌. 响应面法优化枯草芽孢杆菌产脂肪酶的合成培养基[J]. 中国生物工程杂志, 2010, 30(08): 100-105.

SONG Ping, CU Xiao-Ling, HU Yi, XIE Ning-Chang. Optimization of Lipase Production Conditions by Bacillus subtilis Using Surface Methodology. China Biotechnology, 2010, 30(08): 100-105.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/Q815 或 https://manu60.magtech.com.cn/biotech/CN/Y2010/V30/I08/100

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